US4356149A - Multi-layer chemical analytical materials - Google Patents

Multi-layer chemical analytical materials Download PDF

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US4356149A
US4356149A US06/165,444 US16544480A US4356149A US 4356149 A US4356149 A US 4356149A US 16544480 A US16544480 A US 16544480A US 4356149 A US4356149 A US 4356149A
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layer
reagent
hydrophilic
fine particles
water
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Masao Kitajima
Fuminori Arai
Asaji Kondo
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Fujifilm Holdings Corp
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Fuji Photo Film Co Ltd
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/58—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving urea or urease
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/52—Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements
    • G01N33/525—Multi-layer analytical elements
    • G01N33/526—Multi-layer analytical elements the element being adapted for a specific analyte
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S435/00—Chemistry: molecular biology and microbiology
    • Y10S435/805—Test papers

Definitions

  • This invention relates to an improved analytical material for use in the quantitative determination of a specific component contained in an aqueous liquid sample. More particularly, it is concerned with a multi-layer chemical analytical material comprising a support, a reagent layer and a spreading layer which is characterized in that the reagent layer is of an oil-in-water type dispersion wherein the reagent is contained in hydrophobic particles dispersed in a hydrophilic binder.
  • Multi-layer chemical analytical materials in sheet form which permit analysis by a dry operation are described, for example, in Japanese Patent Publication No. 33800/74 (corresponding to U.S. Pat. No. 3,630,957), Japanese Patent Application (OPI) Nos. 53888/74 (corresponding to U.S. Pat. No. 3,992,158), 137192/75 (corresponding to U.S. Pat. No. 3,983,005), 40191/76 (corresponding to U.S. Pat. No. 4,042,335), 3488/77 (corresponding to U.S. Pat. No. 4,006,403), 89796/78 (corresponding to U.S. Pat. No. 4,069,017), 131089/78 (corresponding to U.S. Pat. No. 4,144,306), etc.
  • reagent layer of a single or multi-layer construction is provided on a support and an isotropically porous spreading layer is provided on the reagent layer.
  • an aqueous liquid sample On dropping an aqueous liquid sample on the spreading layer, the aqueous liquid sample penetrates into the reagent layer and undergoes a color-forming reaction, and this change in color density permits determination of the concentration of a certain component in the aqueous liquid sample.
  • urea is subjected to enzyme decomposition with a first reagent, urease, to form ammonia and the ammonia so formed is allowed to participate in the color formation by use of a second reagent, for example, a pH indicator or a mixed reagent of a diazonium salt and a coupler, whereby the quantitative determination of urea can finally be achieved.
  • a second reagent for example, a pH indicator or a mixed reagent of a diazonium salt and a coupler
  • all of the reagents can be incorporated in the reagent layer of the single layer construction whereas for other multi-layer analytical sheets it is preferred that the reagents are divided into the first reagent and the second reagent according to the order of reaction and separately incorporated in a plurality of reagent layers.
  • the multi-layer analytical sheet as described in U.S. Pat. No. 3,011,874 and Japanese Patent Application (OPI) No. 3488/77 is of a complicated multi-layer construction wherein a reagent layer is divided into two layers, one of the layers being composed of a hydraulic binder and the other being composed of a hydrophilic binder or protected by a film of a hydrophobic substance so as to prevent water from penetrating therein. That is, the quantitative determination of a certain component in an aqueous liquid sample is carried out by reaction in water followed by reaction in a non-aqueous medium.
  • a method wherein coating and drying are successively carried out and at least two coating-drying steps are required.
  • the hydrophilic binder layer is provided directly on the hydrophobic layer, the adhesion between the two layers is weak and they often separate from each other in a short time.
  • the hydrophobic binder layer is first provided, at least one additional adhesive layer called an undercoating layer or intermediate layer is then provided on the hydrophobic binder layer, and the hydrophilic binder layer is finally coated on the adhesive layer.
  • the multi-layer analytical sheet of the complicated multi-layer construction in which the reagents are divided into a hydrophilic binder layer and the hydrophobic binder layer.
  • the multi-layer analytical material of this invention is characterized in that the reagent layer is not divided into two layers and a single reagent layer composed of a hydrophilic binder and hydrophobic fine particles dispersed therein containing the reagent is employed.
  • a multi-layer analytical material containing therein a reagent layer prepared by dissolving in an organic solvent a hydrophobic reagent which is insoluble or sparingly soluble in water and which is soluble in an organic solvent, and dispersing in a hydrophilic binder polymer the organic solvent solution of the reagent as fine particles is described in Example 4 of Japanese Patent Application (OPI) No. 26188/78 (corresponding to U.S. Pat. No. 4,089,747) and in Japanese Patent Application (OPI) No. 73096/79 (corresponding to British Patent Publication No. 2 007 360A).
  • the reagent layer of such multi-layer analytical material a complicated procedure is required for dispersing in the hydrophilic binder polymer the reagent which is insoluble or sparingly soluble in water such that the reagent which is insoluble or sparingly soluble in water is first dissolved in a solution of the organic solvent to prepare a mixed solution which is then dispersed as oily liquid fine particles.
  • fine particles containing the reagent functions not only to disperse in the hydrophilic binder polymer but also to prevent the permeation of water from the surface portion into the inside portion of the fine particles containing the reagent.
  • This invention provides:
  • a multi-layer chemical analytical material comprising a light-transmitting and water-impermeable support, a reagent layer on the support and an aqueous liquid sample-spreading layer on the reagent layer wherein the reagent layer is composed of a hydrophilic binder and fine hydrophobic particles dispersed in the hydrophilic binder, the particles containing a regent capable of undergoing a direct or indirect reaction with the component to be analyzed to produce a color change;
  • FIG. 1 is an illustrative cross sectional view of a conventional multi-layer analytical sheet
  • FIG. 2 is an illustrative cross sectional view of an embodiment of the multi-layer chemical analytical material of this invention.
  • One of the advantages of this invention is that the single layer construction of the reagent layer permits the coating step to be simplified to at least 1/2 that of the conventional multi-layer construction method as described previously.
  • the apparent detection limit may be increased.
  • the reason for this is considered that in the multi-layer analytical material of this invention including the single reagent layer in which the hydrophobic fine particles containing the reagent are dispersed, as apparent from the geometric structure, the distance between the reagent contained in the hydrophobic fine particle and the reagent present in the hydrophilic binder is short and the contact area between them is markedly enlarged in comparison with the conventional multi-layer analytical sheet provided with a plurality of reagent layers, which allows various reactions to proceed more smoothly until a color is formed.
  • Light-transmitting and water-impermeable supports for use in the multi-layer analytical material of this invention include plastic films such as polyethylene terephthalate, cellulose esters (e.g., cellulose diacetate, cellulose triacetate and cellulose acetate propionate), polycarbonate and polymethyl methacrylate, and glass plates; known transparent supports with a thickness of from about 50 ⁇ m to about 2 mm can be used.
  • plastic films such as polyethylene terephthalate, cellulose esters (e.g., cellulose diacetate, cellulose triacetate and cellulose acetate propionate), polycarbonate and polymethyl methacrylate, and glass plates; known transparent supports with a thickness of from about 50 ⁇ m to about 2 mm can be used.
  • the support is hydrophobic and the adhesion between the support and the hydrophilic binder of the reagent layer is insufficient, it may be subjected to processings to render the surface hydrophilic (e.g., ultraviolet ray irradiation, electron beam irradiation, flame processing, hydrolysis using alkali, etc.), or auxiliary processings, for example, the formation of an undercoating layer made up of a substance having appropriate adhesion to both the support and the hydrophilic binder of the reagent layer on the surface of the support and the formation of minute unevenness on the surface of the support to such an extent as not to greatly reduce the light transmission property (e.g., brushing and electrolytic etching).
  • processings to render the surface hydrophilic e.g., ultraviolet ray irradiation, electron beam irradiation, flame processing, hydrolysis using alkali, etc.
  • auxiliary processings for example, the formation of an undercoating layer made up of a substance having appropriate adhesion to both the support and
  • the reagent which can be incorporated can be selected depending upon the nature of the component to be determined. However, those described in Japanese Patent Application (OPI) No. 3488/77 (corresponding to U.S. Pat. No. 4,066,403) are preferred as the reagent to be incorporated in the chemical analytical material of the present invention.
  • the hydrophobic fine particles of this invention are liquid or solid minute particles containing therein the reagent, which particles are permeable to a gas or a substance soluble in the hydrophobic solvent through diffusion or extraction, but not by water.
  • Such hydrophobic fine particles containing the reagent can be prepared by the same method as in Japanese Patent Application (OPI) No. 13320/74 (corresponding to U.S. Pat. No. 3,951,851) and exemplify oily minute particles prepared by adding a lipophilic binder polymer, tackifier, dye or pigment to oily matters prepared by dissolving the reagent in an organic solvent substantially incompatible with water; oily minute particles prepared by dissolving in an organic solvent substantially incompatible with water, a polymer soluble in an organic solvent, and dissolving or dispersing the reagent in the resulting solution; minute particles prepared by pulverizing a solid polymer containing the reagent; etc.
  • OPI Japanese Patent Application
  • microcapsules prepared by encapsulating the above oily minute particles or minute particles prepared by pulverizing the solid polymer containing the reagent with a thin layer of a lipophilic polymer or a hydrophilic polymer can be used.
  • Organic solvents substantially incompatible with water which are used for the preparation of the minute particles as described above include liquid plasticizers such as phthalic acid esters (e.g., dimethyl phthalate, diethyl phthalate, dibutyl phthalate, dicyclohexyl phthalate and dioctyl phthalate), adipic acid esters (e.g., diisodecyl adipate and dioctyl adipate), phosphoric acid esters (e.g., triethyl phosphate, tributyl phosphate, trioctyl phosphate and triphenyl phosphate); organic solvents such as toluene, xylene, mesitylene, alkylnaphthalenes, methyl acetate, ethyl acetate and butyl acetate; animal oils; vegetable oils and mineral oils.
  • liquid plasticizers such as phthalic acid esters (e.g., dimethyl phthalate,
  • Organic solvent-soluble polymers which are used for the preparation of the minute particles such as described above include cellulose diacetate, cellulose triacetate, cellulose acetate propionate, ethyl cellulose, methyl methacrylate, polystyrene and polycarbonate of disphenol A.
  • the organic solvent forms a solution
  • the reagent is dissolved or dispersed in the resulting solution
  • the solution or dispersion so obtained is added to an aqueous solution of a hydrophilic binder and emulsified or dispersed, and then a part or all of the organic solvent and water are removed by evaporation whereby there can be formed the reagent layer wherein the fine particles containing therein the reagent are dispersed in the hydrophilic binder.
  • microcapsules containing therein the reagent prepared by the same method as described in Japanese Patent Application (OPI) No. 13320/74 (corresponding to U.S. Pat. No. 3,951,851) can also be used as the fine particles.
  • the thickness of the reagent layer is from about 0.5 ⁇ m to about 50 ⁇ m and preferably from about 1 ⁇ m to about 30 ⁇ m.
  • the size of the hydrophobic fine particles containing the reagent is from about 0.01 ⁇ m to about 20 ⁇ m and preferably from about 0.1 ⁇ m to about 10 ⁇ m.
  • Each particle contains the major portion of the reagent in the inner portion thereof, exposing substantially no reagents on the surface thereof and does not allow water to permeate therein.
  • the amount of the hydrophobic fine particles containing the reagent is from about 3% to about 90% and preferably about 5% to about 80% based on the total weight of the reagent layer.
  • Hydrophilic binders for use in the reagent layer include water-soluble proteins such as gelatin, albumin and collagen; vegetable gums such as agar, sodium alginate and agarose; and water-soluble synthetic polymers such as an olefin-maleic anhydride copolymer, polyvinyl alcohol, polyvinyl pyrrolidone, polyacrylamide and poly(sodium benzenesulfonate). These binders are able to form a film containing the reagent and the dry film so formed readily allows an aqueous solution containing a component to be determined (aqueous liquid sample) to permeate therein.
  • a reagent which reacts with the component to be determined, e.g., urease.
  • nonfibrous isotropically porous materials are those wherein fine porous particles of a brush polymer (generally called a membrane filter), diatomaceous earth or fine crystalline materials (e.g., fine crystalline cellulose) are uniformly dispersed in the binder; porous materials wherein fine spherical beads of glass or a synthetic polymer are brought in point contact with each other and held there with a binder; and a brush polymer in which fine powders of TiO 2 , BaSO 4 or the like are uniformly dispersed.
  • a brush polymer generally called a membrane filter
  • diatomaceous earth or fine crystalline materials e.g., fine crystalline cellulose
  • fine crystalline materials e.g., fine crystalline cellulose
  • Such fabrics which are made hydrophilic are a fabric which is degreased by fully washing with water and then dried; and a fabric which is degreased by fully washing with water and then impregnated with a small amount of a hydrophilic polymer, a surfactant, a wetting agent, or a hydrophilic polymer having TiO 2 or BaSO 4 fine powders dispersed therein.
  • a hydrophilic fabric as the aqueous liquid sample-spreading layer and the details of such fabrics are described in Japanese Patent Application No. 72047/79, filed June 8, 1979 (corresponding to U.S. Pat. No. 4,292,272.
  • the aqueous liquid sample-spreading layer is made up of the nonfibrous isotropically porous material
  • its thickness is from about 50 ⁇ m to about 500 ⁇ m, preferably from about 80 ⁇ m to about 300 ⁇ m.
  • the thickness of the fabric which has been made hydrophilic and allowed to dry is from about 80 ⁇ m to about 1 mm, preferably from about 100 ⁇ m to about 400 ⁇ m.
  • the aqueous liquid sample-spreading layer made up of the nonfibrous isotropically porous material can be made as described in Japanese Patent Application (OPI) Nos. 53888/74 (corresponding to U.S. Pat. No. 3,992,158), 137192/75 (corresponding to U.S. Pat. No. 3,983,005), etc.
  • a solution or dispersion capable of forming a nonfibrous isotropically porous layer is coated on a reagent layer and dried, or a thin layer of the nonfibrous isotropically porous material is bonded to the reagent layer.
  • the fabric can be bonded with the reagent layer.
  • the nonfibrous isotropically porous material or the fabric which has been made hydrophilic onto the reagent layer
  • a method in which while the reagent layer is wet or after wetting the dry surface of the reagent layer with water or water containing surfactant, the nonfibrous isotropically porous material or the fabric which has been made hydrophilic is brought in close contact with the reagent layer and bonded, if desired, by applying an appropriate pressure, by utilizing the characteristics of the hydrophilic binder polymer contained in the reagent layer.
  • Another method utilizes an adhesive which is permeable in the aqueous liquid sample.
  • an adhesive layer permeable in the aqueous liquid sample is provided on the reagent layer.
  • a second reagent layer see U.S. Pat. No. 3,992,158
  • a detecting layer see U.S. Pat. No. 4,042,335
  • a barrier layer see U.S. Pat. Nos. 3,983,005 and 4,066,403
  • a radiation-blocking layer or a light reflection layer can be provided between the reagent layer and the aqueous liquid sample-spreading layer. Additionally, between the aqueous liquid sample-spreading layer and the reagent layer or the radiation-blocking layer or the light reflection layer can be provided an adhesive layer for which the aqueous liquid sample is permeable, for the purpose of firmly bonding the aqueous liquid sample-spreading layer.
  • the radiation-blocking layer or light reflection layer and adhesive layer are described in detail in the patent specifications as described hereinbefore (e.g., U.S. Pat. Nos. 3,992,158 and 4,042,335). Referring to these descriptions they can be provided in this invention.
  • a hydrophilic binder polymer and a fine white powder such as a TiO 2 fine powder and a BaSO
  • a layer made up of the same polymer as the aqueous liquid sample-permeable hydrophilic polymer used as a binder in the reagent layer, radiation-blocking layer or light reflection layer and having a thickness of from about 0.5 ⁇ m to about 10 ⁇ m, preferably from about 0.7 ⁇ m to about 5 ⁇ m can be used.
  • the aqueous solution of the hydrophilic polymer is coated on the reagent layer, radiation-blocking layer or light reflection layer and then while it is still wet or after it is dried, the surface is wetted with water or water containing a surfactant, the nonfibrous isotropically porous material or the fabric which has been made hydrophilic is brought into contact with the surface of the adhesive layer and bonded by applying an appropriate pressure.
  • a solution or dispersion capable of forming a nonfibrous isotropically porous layer may be coated on the adhesive layer to obtain the multi-layer chemical analytical material wherein the aqueous liquid sample-spreading layer is firmly bonded.
  • FIG. 1 is an illustrative view of a conventional multi-layer analytical sheet for use in the quantitative determination of the amount of urea in serum
  • an ammonia gas-permeable and water-impermeable hydrophobic second reagent layer 22 composed of a hydrophobic binder and an indicator contained therein, the indicator undergoing discoloration by the action of ammonia gas is provided on a water-impermeable transparent support 1, and a hydrophilic first reagent layer 21 composed of gelatin and urease and a buffering agent dispersed therein is provided on the hydrophobic second reagent layer 22, and furthermore a nonfibrous isotropically porous spreading layer 3 for spreading an aqueous liquid sample is provided on the first reagent layer 21.
  • the serum On attaching a droplet of serum on the spreading layer 3 from the direction of A, the serum uniformly extends so that the volume per unit area is nearly uniform and reaches the first reagent layer 21 where the urea is decomposed by urease.
  • the ammonia so formed is gasified and permeates the second reagent layer 22 where the pH indicator undergoes discoloration depending upon the amount of ammonia.
  • the degree of discoloration is examined from the direction of B and the amount of urea is quantitatively determined.
  • FIG. 2 is an illustrative view of an embodiment of the multi-layer analytical material of this invention wherein a single reagent layer 23 composed of a hydrophilic binder and hydrophobic fine particles 20 dispersed therein is provided on a light-transmitting and water-impermeable support 1, and porous spreading layer 4 is provided on the reagent layer 23.
  • a hydrophobic oil or hydrophobic binder containing the indicator is emulsified and dispersed in a hydrophilic binder aqueous solution containing an enzyme system to give a dispersion and the dispersion so prepared is coated to provide the reagent layer 23.
  • the determination of urea in serum is carried out in the same manner as explained for the conventional one of FIG. 1.
  • the multi-layer analytical material of this invention is advantageous in its ease of production in comparison with the conventional analytical sheet.
  • the hydrophobic reagent solution was emulsified and dispersed in the hydrophilic reagent solution by a conventional method using a supersonic emulsifier to form an oil-in-water (O/W) type emulsion.
  • the size of the hydrophobic particles containing the bromothymol blue was 5 ⁇ m or less.
  • the dispersion thus obtained was coated on a 170 ⁇ m thick transparent polyethylene terephthalate (PET) film which had been undercoated to form a reagent layer.
  • PET transparent polyethylene terephthalate
  • the thickness of the layer after drying was about 20 ⁇ m, and microscopic analysis confirmed that the reagent layer was made up of fine oil particles dispersed in the gelatin phase.
  • a membrane filter (Microfilter FM 500 produced by Fuji Photo Film Co., Ltd.) was laminated on and bonded together with the reagent layer by pressing under humid conditions to provide thereon a porous spreading layer. A multi-layer chemical analytical material for the quantitative determination of an urea aqueous solution was thus obtained.
  • the calibration curve obtained from the above values is nearly a straight line. Therefore, it can be seen that the urea content of an urea aqueous solution can be quantitatively determined by use of the multi-layer chemical analytical material as obtained above.
  • the hydrophobic reagent-polymer solution was added to the hydrophilic reagent-binder aqueous solution, and the hydrophobic reagent-polymer solution was emulsified and dispersed in the hydrophilic-binder aqueous solution by a conventional method using a supersonic emulsifier.
  • the dispersion was stirred for 30 minutes at 55° C. whereby the organic solvent (methyl acetate) was evaporated off to prepare a dispersion having solid particles of ethyl cellulose (polymer) containing bromothymol blue (pH indicator) dispersed in the hydrophilic reagent-binder aqueous solution.
  • the size of the solid particles of the polymer containing bromothymol blue was about 5 ⁇ m or less.
  • the dispersion thus obtained was coated on a 170 ⁇ m thick transparent PET film which has been undercoated with gelatin, and then dried to form a reagent layer in which the solid fine particles of the polymer were dispersed.
  • the thickness of the reagent layer after drying was about 20 ⁇ m, and microscopic analysis confirmed that the reagent layer was made up of the solid fine particles of the polymer dispersed in gelatin.
  • a membrane filter (Microfilter FM 500 produced by Fuji Photo Film Co., Ltd.) was laminated on and bonded together with the reagent layer by pressing under humid conditions to provide thereon a porous spreading layer. A multi-layer chemical analytical material for the quantitative determination of serum urea was thus obtained.
  • Each of buffer aqueous solutions having different pH values ranging between 6 and 8 was applied onto the porous spreading layer of the thus obtained multi-layer chemical analytical material, and then allowed to stand at 37° C. for 5 minutes. No color change of the pH indicator in the reagent layer of each analytical material was observed. This indicates that the solid fine particles of the polymer containing the pH indicator in the multi-layer chemical analytical material were water-impermeable and that the pH indicator (bromothymol blue) was not present in a substantially exposed state on the surfaces of the solid fine particles of the polymer.
  • the calibration curve obtained from the above values is nearly a straight line. Therefore, it can be seen that the urea content in the serum can be quantitatively determined by use of the multi-layer chemical analytical material as obtained above.
  • a hydrophilic binder solution 20 g of a 10% by weight aqueous solution of gelatin was prepared.
  • the hydrophobic reagent solution was emulsified and dispersed in the hydrophilic reagent solution by a conventional method and by continuing the stirring at 50° C.
  • the solvents ethyl acetate and toluene
  • the solvents were evaporated to produce a dispersion wherein microcapsules in which hydrophobic solid particles containing the indicator were encapsulated with a thin film of ethyl cellulose were dispersed in the hydrophilic binder (gelatin) aqueous solution.
  • Example 2 The thus obtained solution was coated on a transparent PET film and dried in the same manner as in Example 1, and subsequently the same porous spreading layer as described in Example 1 was laminated thereon to provide a multi-layer chemical analytical material.
  • each of buffer aqueous solutions having different pH values ranging between 6 and 8 was applied onto the porous spreading layer of the thus obtained multi-layer chemical analytical material. No color change of the pH indicator was observed. This indicates that the hydrophobic fine particles (microcapsules) containing the pH indicator in the multi-layer chemical analytical material were water-impermeable.
  • Example 2 To this solution was further added a solution prepared by dissolving 20 mg of urease and 200 mg EDTA 2Na in 2 ml of water. The resulting solution was coated on a transparent PET film and dried in the same manner as in Example 1 to provide an about 15 ⁇ m thick reagent layer containing hydrophobic fine particles (microcapsules) which contained the reagent.
  • Example 2 On the reagent layer was coated a 5% aqueous solution of gelatin containing 35% of fine titanium dioxide particles to provide thereon a 17 ⁇ m thick radiation-blocking layer. Furthermore, on the radiation-blocking layer was laminated the same porous spreading layer as in Example 1 to provide a multi-layer chemical analytical material.
  • Synthetic serums with different concentrations of urea were prepared by adding urea to a phosphoric acid buffer aqueous solution containing 7% of serum protein and having a pH value of 7.0. These serums were tested on the multi-layer chemical analytical material as obtained above, in the same manner as in Example 1. The results are shown in the table below.
  • One surface of a broadcloth (a product of Nisshin Spinning Co., Ltd.) woven at a No. 60 cotton count yarn was made hydrophilic by impregnating it with a 1% aqueous solution of gelatin to prepare a fabric having a gelatin content of about 2.5% at drying.
  • a reagent layer and a radiation-blocking layer were provided on a transparent PET film and dried in the same manners as those employed in Example 1 and Example 3, respectively.
  • the radiation-blocking layer was wetted and swollen with a 0.2% aqueous solution of a nonionic surface active agent (polyoxyethylene isooctylphenyl ether), and the above-described fabric which had been made hydrophilic was laminated on and bonded together with the thus treated radiation-blocking layer by lightly pressing in such a manner that the surface of the fabric which had been impregnated with the gelatin aqueous solution was brought into contact with the radiation-blocking layer.
  • the fabric which had been made hydrophilic functions as the spreading layer for the quantitative determination of an aqueous liquid sample.
  • Quantitative determination of blood urea could be made in the same manner as in Example 1 using fresh blood (whole blood) by the use of the thus prepared multi-layer chemical analytical material for the quantitiative analysis of blood urea.
  • Example 4 The same procedure as in Example 2 were repeated except that the cotton fabric prepared in the same manner as in Example 4 was used in place of the membrane filter and laminated on the radiation-blocking layer in the same manner as in Example 4, whereby a multi-layer chemical analytical material was prepared.
  • Example 4 The same procedures as in Example 3 were repeated except that the cotton fabric prepared in the same manner as in Example 4 was used in place of the membrane filter and laminated on the radiation-blocking layer in the same manner as in Example 4, whereby a multi-layer chemical analytical material was prepared.
  • Example 3 The test was carried out in the same manner as in Example 3 using buffer aqueous solutions having different pH values ranging between 6 and 8 and synthetic serums whereby the same results as in Example 3 were obtained.

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Cited By (55)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE3235658A1 (de) * 1981-09-29 1983-04-14 Konishiroku Photo Industry Co., Ltd., Tokyo Analyseeinheit
US4427632A (en) 1981-04-29 1984-01-24 Konishiroku Photo Industry Co., Ltd. Analytical element
EP0103288A1 (en) 1982-09-10 1984-03-21 Fuji Photo Film Co., Ltd. Multilayer analytical element
EP0141647A3 (en) * 1983-11-07 1985-06-19 Technicon Instruments Corporation Reflective particle-containing analysis composition and device
JPS60115863A (ja) * 1983-11-07 1985-06-22 テクニコン、インストルメンツ、コーポレーシヨン 反射性粒子含有分析用素子
US4549655A (en) * 1983-12-05 1985-10-29 E. I. Du Pont De Nemours And Company Container for a layered chemical analysis system
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US20030017464A1 (en) * 2001-07-17 2003-01-23 Ciphergen Biosystems, Inc. Latex based adsorbent chip
US20030113932A1 (en) * 2001-12-14 2003-06-19 Shmuel Sternberg Hydrophobic ammonia sensing membrane
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US20050129572A1 (en) * 2002-01-15 2005-06-16 Schulman Lloyd S. Liquid permeable composition in dry reagent devices
US20050180879A1 (en) * 2004-02-03 2005-08-18 Eva Hrboticka One step tester for ammonia
US20050287682A1 (en) * 2004-06-28 2005-12-29 Lizzi Michael J Dissolvable films and methods including the same
WO2007017260A3 (de) * 2005-08-11 2007-05-18 Univ Albert Ludwigs Freiburg Sensor mit mehrphasigem, segmentiertem polymernetzwerk
US20090053738A1 (en) * 2006-02-23 2009-02-26 Mologic Ltd. Protease detection
US20090098020A1 (en) * 2006-02-23 2009-04-16 Mologic Ltd Binding Assay
US20090123955A1 (en) * 2004-12-13 2009-05-14 Bayer Healthcare Llc Size Self-Limiting Compositions and Test Devices for Measuring Analytes in Biological Fluids
US20090191582A1 (en) * 2006-02-23 2009-07-30 Mologic Ltd Enzyme detection
US20090203059A1 (en) * 2006-04-07 2009-08-13 Mologic Ltd. Protease detection product
WO2012108907A1 (en) * 2011-02-11 2012-08-16 Ansell Limited Breach or contamination indicating article having microcapsules
US20150004707A1 (en) * 2013-06-26 2015-01-01 Mridula Nair Methods for using indicator compositions
US20150004706A1 (en) * 2013-06-26 2015-01-01 Mridula Nair Reactive indicator compositions and articles containing same
EP2969566A4 (en) * 2013-03-12 2016-11-23 Ansell Ltd ARTICLE INDICATOR OF PENETRATION OR CONTAMINATION
US10065210B2 (en) 2014-06-12 2018-09-04 Ansell Limited Breach or contamination indicating article, optionally with pre-warning indicator
DE102017128564A1 (de) * 2017-12-01 2019-06-06 Endress+Hauser Conducta Gmbh+Co. Kg Sensormembran, Sensorkappe, optischer Sensor und Verfahren zur Herstellung einer Sensormembran
WO2020219130A1 (en) 2019-04-24 2020-10-29 Progenitec, Inc. System for analysis of body fluids and wound-associated biomolecules
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JPS6014141A (ja) * 1983-07-06 1985-01-24 Kyoto Daiichi Kagaku:Kk 多層一体化分析用具
JPH01197654A (ja) * 1988-02-02 1989-08-09 Eiken Kagaku Kk 乾式分析材料
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US4615983A (en) * 1980-11-11 1986-10-07 Konishiroku Photo Industry Co., Ltd. Immunological measuring element
US4427632A (en) 1981-04-29 1984-01-24 Konishiroku Photo Industry Co., Ltd. Analytical element
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DE3235658A1 (de) * 1981-09-29 1983-04-14 Konishiroku Photo Industry Co., Ltd., Tokyo Analyseeinheit
EP0103288A1 (en) 1982-09-10 1984-03-21 Fuji Photo Film Co., Ltd. Multilayer analytical element
US4557900A (en) * 1982-09-28 1985-12-10 Cardiovascular Devices, Inc. Optical sensor with beads
US4803158A (en) * 1983-03-08 1989-02-07 Kabushiki Kaisha Kyoto Daiichi Kagaku Composition used for the determination of beta-hydroxybutyric acid and a method for preparing the said composition
US4649123A (en) * 1983-05-12 1987-03-10 Miles Laboratories, Inc. Ion test means having a hydrophilic carrier matrix
US4729948A (en) * 1983-09-12 1988-03-08 Konishiroku Photo Industry Co., Ltd. Dry analytical element for α-amylase
EP0141647A3 (en) * 1983-11-07 1985-06-19 Technicon Instruments Corporation Reflective particle-containing analysis composition and device
JPS60115863A (ja) * 1983-11-07 1985-06-22 テクニコン、インストルメンツ、コーポレーシヨン 反射性粒子含有分析用素子
JPS60115862A (ja) * 1983-11-07 1985-06-22 テクニコン、インストルメンツ、コーポレーシヨン 反射性粒子含有溶媒抽出試薬組成物
US4549655A (en) * 1983-12-05 1985-10-29 E. I. Du Pont De Nemours And Company Container for a layered chemical analysis system
US4670218A (en) * 1984-02-24 1987-06-02 Miles Laboratories, Inc. Ion test means having a porous carrier matrix
EP0175195A3 (en) * 1984-09-13 1988-10-05 Behringwerke Aktiengesellschaft Device comprising membrane-structured polymers with solid particles incorporated there-in
EP0175990A3 (en) * 1984-09-22 1988-09-21 Bayer Ag Membrane for carrying reagents, method for its preparation and its use as an analytical means and in analytical methods
US4780411A (en) * 1984-09-22 1988-10-25 Bayer Aktiengesellschaft Water-absorbing, essentially water-free membrane for reagent substrates and methods of preparing the same
EP0207392A2 (en) 1985-07-02 1987-01-07 Miles Inc. Multilayer ion test means
EP0212989A3 (en) * 1985-08-30 1988-06-01 Eastman Kodak Company Lipid vesicles containing labeled species and their analytical uses
US5063153A (en) * 1986-10-09 1991-11-05 Fuji Photo Film Co., Ltd. Integral multilayer analytical element
US4789526A (en) * 1986-12-15 1988-12-06 Pall Corporation Vacuum diagnostic device
US4976926A (en) * 1986-12-15 1990-12-11 Pall Corporation Vacuum diagnostic device
US5122451A (en) * 1987-12-18 1992-06-16 Fuji Photo Film Ltd. Dry liquid analysis element
US4956146A (en) * 1988-03-14 1990-09-11 Fuji Photo Film Co., Ltd. Dry analytical element and process for producing the same
US5215712A (en) * 1989-09-14 1993-06-01 Terumo Kabushiki Kaisha Apparatus for determination of ion concentration, specific gravity, or osmotic pressure of solution
US5322797A (en) * 1990-06-06 1994-06-21 Southwest Research Institute Method for detecting vapor and liquid reactants
US5183763A (en) * 1990-06-06 1993-02-02 Southwest Research Institute Composition and method for detecting vapor and liquid reactants
EP0607209A4 (en) * 1991-10-11 1997-01-22 Abbott Lab Unit-of-use reagent compositions for specific binding assays.
US6190612B1 (en) 1998-01-21 2001-02-20 Bayer Corporation Oxygen sensing membranes and methods of making same
US6254831B1 (en) 1998-01-21 2001-07-03 Bayer Corporation Optical sensors with reflective materials
US6306347B1 (en) 1998-01-21 2001-10-23 Bayer Corporation Optical sensor and method of operation
US6387709B1 (en) 1998-01-21 2002-05-14 Bayer Corporation Method of operating an optical sensor adapted for selective analyte-sensing contact with a plurality of samples
US6488891B2 (en) 1998-01-21 2002-12-03 Bayer Corporation Optical sensor and method of operation
US6446516B1 (en) 1998-05-13 2002-09-10 Bayer Corporation Sample introduction device
US6812035B1 (en) 1998-07-10 2004-11-02 Chemmotif, Inc. Dye desortion molecular indicator
EP1097366A4 (en) * 1998-07-10 2002-09-04 Chemmotif Inc MOLECULAR INDICATOR OF DYE DESORPTION
US6107083A (en) * 1998-08-21 2000-08-22 Bayer Corporation Optical oxidative enzyme-based sensors
US20040047768A1 (en) * 1999-05-12 2004-03-11 Sullivan Kevin J. Sample introduction device
US7157056B2 (en) 1999-05-12 2007-01-02 Bayer Corporation Sample introduction device
WO2002103334A1 (de) * 2001-06-18 2002-12-27 Presens Precision Sensing Gmbh Sauerstoffsensoren auf mikrotiterplatte
US20040171094A1 (en) * 2001-06-18 2004-09-02 Ingo Klimant Oxygen sensors disposed on a microtiter plate
US20030017464A1 (en) * 2001-07-17 2003-01-23 Ciphergen Biosystems, Inc. Latex based adsorbent chip
US7517496B2 (en) * 2001-07-17 2009-04-14 Bio-Rad Laboratories, Inc. Latex based adsorbent chip
WO2003079402A3 (en) * 2001-07-17 2004-05-27 Ciphergen Biosystems Inc Latex based adsorbent chip
US20030113932A1 (en) * 2001-12-14 2003-06-19 Shmuel Sternberg Hydrophobic ammonia sensing membrane
US20050129572A1 (en) * 2002-01-15 2005-06-16 Schulman Lloyd S. Liquid permeable composition in dry reagent devices
US20030215358A1 (en) * 2002-01-15 2003-11-20 Schulman Lloyd S. Liquid permeable composition in dry reagent devices
WO2003060517A3 (en) * 2002-01-15 2003-12-24 Bayer Healthcare Llc Diffusable adhesive composition for multi-layered dry reagent devices
US7713474B2 (en) 2002-01-15 2010-05-11 Siemens Healthcare Diagnostics Inc. Liquid permeable composition in dry reagent devices
US20050180879A1 (en) * 2004-02-03 2005-08-18 Eva Hrboticka One step tester for ammonia
US20050287682A1 (en) * 2004-06-28 2005-12-29 Lizzi Michael J Dissolvable films and methods including the same
US9410185B2 (en) 2004-06-28 2016-08-09 Becton, Dickinson And Company Dissolvable films and methods including the same
US9267167B2 (en) * 2004-06-28 2016-02-23 Becton, Dickinson And Company Dissolvable films and methods including the same
US7964372B2 (en) 2004-12-13 2011-06-21 Bayer Healthcare Llc Size self-limiting compositions and test devices for measuring analytes in biological fluids
US20090123955A1 (en) * 2004-12-13 2009-05-14 Bayer Healthcare Llc Size Self-Limiting Compositions and Test Devices for Measuring Analytes in Biological Fluids
US9982289B2 (en) 2004-12-13 2018-05-29 Ascensia Diabetes Care Holdings Ag Size self-limiting compositions and test devices for measuring analytes in biological fluids
WO2007017260A3 (de) * 2005-08-11 2007-05-18 Univ Albert Ludwigs Freiburg Sensor mit mehrphasigem, segmentiertem polymernetzwerk
US20090098020A1 (en) * 2006-02-23 2009-04-16 Mologic Ltd Binding Assay
US7935497B2 (en) 2006-02-23 2011-05-03 Mologic Ltd Protease detection
US8241588B2 (en) 2006-02-23 2012-08-14 Mologic Ltd Binding assay
US8361386B2 (en) 2006-02-23 2013-01-29 Mologic Ltd Enzyme detection
US8846328B2 (en) 2006-02-23 2014-09-30 Mologic Ltd Method for detecting the presence of a protease enzyme in a sample
US20090191582A1 (en) * 2006-02-23 2009-07-30 Mologic Ltd Enzyme detection
US20090053738A1 (en) * 2006-02-23 2009-02-26 Mologic Ltd. Protease detection
US20090203059A1 (en) * 2006-04-07 2009-08-13 Mologic Ltd. Protease detection product
WO2012108907A1 (en) * 2011-02-11 2012-08-16 Ansell Limited Breach or contamination indicating article having microcapsules
EP2969566A4 (en) * 2013-03-12 2016-11-23 Ansell Ltd ARTICLE INDICATOR OF PENETRATION OR CONTAMINATION
US9579532B2 (en) 2013-03-12 2017-02-28 Ansell Limited Breach or contamination indicating article
US9291570B2 (en) * 2013-06-26 2016-03-22 Eastman Kodak Company Reactive indicator compositions and articles containing same
US9289528B2 (en) * 2013-06-26 2016-03-22 Eastman Kodak Company Methods for using indicator compositions
US20150004706A1 (en) * 2013-06-26 2015-01-01 Mridula Nair Reactive indicator compositions and articles containing same
US20150004707A1 (en) * 2013-06-26 2015-01-01 Mridula Nair Methods for using indicator compositions
US10065210B2 (en) 2014-06-12 2018-09-04 Ansell Limited Breach or contamination indicating article, optionally with pre-warning indicator
DE102017128564A1 (de) * 2017-12-01 2019-06-06 Endress+Hauser Conducta Gmbh+Co. Kg Sensormembran, Sensorkappe, optischer Sensor und Verfahren zur Herstellung einer Sensormembran
WO2020219130A1 (en) 2019-04-24 2020-10-29 Progenitec, Inc. System for analysis of body fluids and wound-associated biomolecules
EP3873394A4 (en) * 2019-04-24 2022-08-17 Progenitec, Inc. SYSTEM FOR ANALYZING BODY FLUIDS AND WOUND BIOMOOLECLES
US11506658B2 (en) 2019-04-24 2022-11-22 Progenitec, Inc. System for analysis of body fluids and wound-associated biomolecules
US11680181B1 (en) 2021-09-29 2023-06-20 W. Thomas Forlander Industrial coating with color-change responsivity to acid and base contact

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JPS6226423B2 (enrdf_load_stackoverflow) 1987-06-09

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